DNA Transformation to PEG
1. Transfer 100µl of competent cells to a 2059 snap-cap
tube on ice.
2. Add 10µl of ligation product or up to 0.5µgr
of super-coiled plasmid.
3. Incubate 30’ at 4oC (on ice)
4. Incubate 45 sec in 42 oC *
do not exceed 45''
5. Incubate 2 min on ice
6. Add 0.9 ml of Room Temp. SOC medium to the
7. Incubate in shaker at 37oC for 1 hour (up
to 90min) * The purpose of this step is to let the
bacteria recover from the heat shock prior to plating on agar plates. Thus
no antibiotic is added to the medium during this step, and the medium should
not be cold.
8. Seed on plates 100µl of bugs , then spin the
bugs 30'', aspirate sup and leave about 100µl, re-suspend the bugs
in the medium, and plate all. * make sure you add
the right antibiotics to the agar plates - check vector AND bacterial